Detection and quantitation of HIV has been a longstanding issue for scientists and clinicians since the discovery of the virus in 1984. Currentl there are several assays available for screening for HIV infection in the field or the laboratory. They typically fall into a few categories: PCR-based assays that detect HIV RNA, ELISA assays that detect p24 capsid protein, and assays that detect HIV reverse transcriptase (RT). There are also rapid detection screening assays that detect antibodies to HIV and are typically used as the initial screening tests in field assays because of they are highly sensitive and take just minutes to complete. The RT assays though cost effective in most cases, tend to have low sensitivity which is also a drawback of p24 assays. PCR-based assays have high sensitivity but are typically costly and time consuming and require specialized equipment. These assays are also subject to contamination like other PCR assays. The HIV field would benefit from an inexpensive assay that is sensitive, quantitative, fast and simple. An assay that can be performed in a laboratory setting with small quantities of virus and could detect even very low levels of infection that might result from drug screens or mutational analysis. In this application we propose such an assay using aptamer-based technology that we have developed in our lab. Our lab has developed nucleic acid aptamers that can bind to HIV RT with very tightly and are capable of detecting as few as 50-100 virus particles. We have used these aptamers to begin to develop an Aptamer-based RT detection Assay (ARTA) that is sensitive, quantitative, and inexpensive. We envision this technology as the next generation of HIV RT-based detection assays and as a model for development of other aptamer-based pathogen detection assays.
This work is relevant to human health because it will develop a new assay that can detect and quantify human immunodeficiency virus. The assay is highly sensitive, simple to perform, and has low cost.
|DeStefano, Jeffrey J; Alves Ferreira-Bravo, Irani (2018) A highly sensitive aptamer-based HIV reverse transcriptase detection assay. J Virol Methods 257:22-28|
|Alves Ferreira-Bravo, Irani; Cozens, Christopher; Holliger, Philipp et al. (2015) Selection of 2'-deoxy-2'-fluoroarabinonucleotide (FANA) aptamers that bind HIV-1 reverse transcriptase with picomolar affinity. Nucleic Acids Res 43:9587-99|